State Key Lab of Rice Biology, China National Rice Research Institute, Hangzhou, 310006, Zhejiang, China.
College of Biotechnology and Biongineering, Zhejiang University of Technology, Chaowang Road, Hangzhou, 310014, Zhejiang, China.
Sci Rep. 2020 Jul 2;10(1):10885. doi: 10.1038/s41598-020-67735-7.
Ten distinct cDNAs encoding five different protein phosphatases 1 (PPP1) were cloned from Nilaparvata lugens. NlPPP1α and NlPPP1β are highly conserved whereas NlPPP1-Y, NlPPP1-Y1 and NlPPP1-Y2 are lowly conserved among insects. NlPPP1α and NlPPP1β exhibited a ubiquitous expression, while NlPPP1-Y, NlPPP1-Y1, and NlPPP1-Y2 were obviously detected from the 4th instar nymph to imago developmental stages in males, especially detected in internal reproductive organ and fat bodies of the male. Injection nymphs with dsRNA of NlPPP1α or NlPPP1β was able to reduce the target gene expression in a range of 71.5-91.0%, inducing a maximum mortality rate of 95.2% or 97.2% at 10th day after injection and eclosion ratio down by 65.5-100.0%. Injection with dsNlPPP1Ys targeted to NlPPP1-Y, NlPPP1-Y1and NlPPP1-Y2 was able to induce a maximum mortality rate of 95.5% at 10th day after injection, eclosion ratio down by 86.4%. Knock-down one of the male-biased NlPPP1 genes has no effect on survival and eclosion ratio. Injection of 4th instar nymph with dsNlPPP1Ys led to reduced oviposition amount and hatchability, down by 44.7% and 19.6% respectively. Knock-down of NlPPP1-Y1 or NlPPP1-Y2 gene did not significantly affect oviposition amount but significantly affected hatchability. The results indicate that the male-biased NlPPP1 genes have overlapping functions in N. lugens development, and NlPPP1-Y1 and NlPPP1-Y2 may play important roles in spermatogenesis and fertilization. The dsNlPPP1β and dsNlPPP1Ys in this study could be the preferred sequence in RNAi and low-conserved male-biased NlPPP1 genes could be potential target for N. lugens control.
从褐飞虱中克隆了 10 种不同的编码 5 种不同蛋白磷酸酶 1（PPP1）的 cDNA。NlPPP1α 和 NlPPP1β 在昆虫中高度保守，而 NlPPP1-Y、NlPPP1-Y1 和 NlPPP1-Y2 则低度保守。NlPPP1α 和 NlPPP1β 表现出普遍表达，而 NlPPP1-Y、NlPPP1-Y1 和 NlPPP1-Y2 则在雄性 4 龄若虫到成虫发育阶段明显检测到，特别是在雄性的内部生殖器官和脂肪体中检测到。注射 NlPPP1α 或 NlPPP1β 的 dsRNA 能够在 71.5-91.0%的范围内降低靶基因的表达，在注射后第 10 天诱导最高死亡率为 95.2%或 97.2%，孵化率下降 65.5-100.0%。注射靶向 NlPPP1-Y、NlPPP1-Y1 和 NlPPP1-Y2 的 dsNlPPP1Ys 能够在注射后第 10 天诱导最高死亡率 95.5%，孵化率下降 86.4%。敲除一个雄性偏爱的 NlPPP1 基因对生存和孵化率没有影响。注射 4 龄若虫的 dsNlPPP1Ys 导致产卵量和孵化率降低，分别降低了 44.7%和 19.6%。敲除 NlPPP1-Y1 或 NlPPP1-Y2 基因对产卵量没有显著影响，但对孵化率有显著影响。结果表明，雄性偏爱的 NlPPP1 基因在褐飞虱发育中具有重叠功能，NlPPP1-Y1 和 NlPPP1-Y2 可能在精子发生和受精中发挥重要作用。本研究中的 dsNlPPP1β 和 dsNlPPP1Ys 可能是 RNAi 的首选序列，低保守的雄性偏爱的 NlPPP1 基因可能是褐飞虱控制的潜在靶标。
